• 한국임상수의학회지
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• 제40권1호
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• pp.1-7
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• 2023

The growing market for companion animals, combined with their increasing lifespan, has generated an increased interest in companion animal immunity enhancers. Ginsenoside, a saponin component of ginseng and an essential ingredient of red ginseng marc (produced during red ginseng production), is effective in improving immunity. In this experiment, a powder mixture of red ginseng marc and steamed red ginseng extract powder (RGME) was orally administered to dogs for eight weeks. Subsequently, blood samples were collected and tested every four weeks. In addition, canine peripheral blood mononuclear cells (cPBMCs) were stimulated with or without interleukin-2 (IL-2) to evaluate their proliferation and cytokine secretion abilities. Proliferation assay suggests that the administration of RGME effectively enhanced numbers of cPBMCs under IL-2 stimulation. Furthermore, in the RGME group, a significant increase in the concentration of interferon gamma released from cPBMCs under IL-2 stimulation was observed. In conclusion, RGME might be an effective health supplement for improving immunity in dogs.

• Journal of Pharmaceutical Investigation
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• 제32권4호
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• pp.285-290
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• 2002

In order to assay the efficacy of newly synthesized antiviral compounds, in vitro studies of their active intracellular phosphorylated metabolites were established as compared with Zidovudine (ZDV). Antiviral base analogs require intracellular phosphorylation prior to the inhibition of HIV replication. Therefore, antiviral drugs concentrations in plasma have not reflected any direct relationship with activity or toxicity. A method has been developed to measure the concentration of total phosphorylated metabolites inside peripheral blood mononuclear cells using modified commercial radioimmunoassay (RIA). ZDV 5'-monophosphate was synthesized and used as a procedural control for RIA modification. PBMCs were isolated from whole blood and incubated with ZDV for 20 h to allow metabolic phosphorylation. Viable cells were extracted overnight with 60% methanol. After evaporation, the extract was reconstituted in Tris buffer. Samples were split into two fractions, one of which was treated with alkaline phosphatase (AP) to liberate phosphate groups. Concentrations of phosphorylated metabolites were determined by subtracting thε concentration of non-AP-treated fraction from that of the treated fraction. Recovery of phosphorylated ZDV from cell extracts was approximately 90%, and reproducibility was acceptable (coefficients of variation <15% for concentrations${\geq}$0.25 ng/mL). Intracellular concentrations $(0.135{\sim}5.019\;nmole/10^6\;cells)$ followed a nonlinear dose-response relationship over the range $0.015{\sim}2.996mM$ extracellular ZDV, with concentration-dependant saturation.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6527-6531
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• 2012

Purpose: Complement receptor 1 (CR1) is induced by Epstein-Barr virus (EBV) and may be a potential biomarker of nasopharyngeal carcinoma (NPC). We conducted the present study to evaluate the association of CR1 expression with clinicopathological features and prognosis of NPC. Methods: We enrolled 145 NPC patients and 110 controls. Expression levels of CR1 in peripheral blood mononuclear cells (PBMCs) were detected using quantitative real-time PCR and associations with clinicopathological features and prognosis were examined. Results: CR1 levels in the NPC group [3.54 (3.34, 3.79)] were slightly higher than those in the controls [3.33 (3.20, 3.47)] (P<0.001). Increased CR1 expression was associated with histology classification (type III vs. type II, P=0.002), advanced clinical stage (P=0.003), high T stage (P=0.017), and poor overall survival (HR, 4.89; 95% CI, 1.23-19.42; P=0.024). However, there were no statistically significant differences in CR1 expression among N or M stages. Conclusion: These findings indicate that CR1 expression in PBMCs may be a new biomarker for prognosis of NPC and a potential therapeutic target.

• 한국임상수의학회지
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• 제33권4호
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• pp.194-199
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• 2016

The objective of this study was to examine the effect of trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) on the expression of cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX) pathway in lipopolysaccharide (LPS)-stimulated porcine peripheral blood mononuclear cells (PBMCs). t10c12-CLA was treated with different concentrations in culture medium of LPS$na{\ddot{i}}ve$ and LPS-stimulated PBMCs. The mRNA expressions of prostaglandin $E_2$ ($PGE_2$)-synthase, COX-2 and 5-LOX were measured using quantitative real-time PCR. In addition, the production levels of $PGE_2$ and 5-LOX in culture supernatant from PBMCs with or without LPS were assessed by ELISA. In LPS$na{\ddot{i}}ve$ PBMCs, treatment of t10c12-CLA significantly (p < 0.05) increased the mRNA expressions of PGE2 synthase and 5-LOX compared to vehicle control. Expression of COX-2 mRNA did not show significant difference compared to vehicle control by t10c12-CLA treatment in LPS$na{\ddot{i}}ve$ PBMCs. However, the addition of LPS in PBMCs markedly (p < 0.05) increased the mRNA expression of COX-2, $PGE_2$ synthase and 5-LOX, and also significantly (p < 0.05) enhanced the production of $PGE_2$ and 5-LOX relative to LPS$na{\ddot{i}}ve$ PBMCs, respectively. However, the addition of t10c12-CLA significantly (p < 0.01) suppressed the LPS-induced excessive expression of COX-2, $PGE_2$ synthase, and 5-LOX compared to those of PBMCs treated with LPS alone. The production levels of $PGE_2$ and 5-LOX in culture supernatant from LPS-stimulated PBMCs were also significantly (p < 0.05) inhibited by the treatment of t10c12-CLA compared to LPS alone. These results suggested that t10c12-CLA has an anti-inflammatory effect via dual inhibition of COX-2 and 5-LOX with gene expression and production level in LPS-stimulated porcine PBMCs. Therefore, it was thought that t10c12-CLA can attenuate the inflammatory response by down-regulation of eicosanoids production.

• 한국임상수의학회지
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• 제32권4호
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• pp.289-294
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• 2015

Fucoidan은 갈조류로부터 추출되는 황산다당류로 다양한 생리학적 활성을 갖고 있다. 본 연구의 목적은 LPS로 자극한 돼지 말초혈액 단핵구세포(PBMCs)의 NO 생산에 있어 fucoidan의 효과를 검토하고, 이러한 효과가 iNOS의 발현과 AP-1의 활성화와 관련이 있는지를 조사하는데 있다. LPS 무처치 돼지 PBMCs에서 fucoidand의 처리는 NO 생산과 AP-1활성에 대해 효과를 보이지 않았다. 또한 iNOS와 AP-1의 mRNA 발현도 fucoidan 처치에 의해 영향을 받지 않았다. 그러나 LPS로 자극한 PBMCs에서는 NO 생산과 AP-1의 활성 그리고 iNOS와 AP-1의 mRNA 발현이 현저하게 증가하였다. 이와 같은 LPS에 의한 돼지 PBMCs의 NO 생산과 AP-1 활성증가는 fucoidan 첨가에 의해 감소되었다. 또한 fucoidan은 LPS에 의한 iNOS와 AP-1의 mRNA 발현증가도 억제시켰다. 이상의 결과는 fucoidan이 LPS 자극 돼지 PBMCs에서 iNOS 발현과 AP-1 활성의 억제와 함께 NO 생산을 하향 조절함으로써 항염증효과를 나타내는 것으로 사료되었다.

• Journal of Animal Science and Technology
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• 제63권6호
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• pp.1376-1385
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• 2021

Foot-and-mouth disease, one of the most contagious diseases in cloven-hoofed animals, causes significant economic losses. The pathogenesis of foot-and-mouth disease virus (FMDV) infection is known to differ with age of the animals. In this study, we aimed to reveal the difference in immunological response in the initial stage of FMDV infection between piglets and adult pigs. Peripheral blood mononuclear cells (PBMCs) were isolated from 3 piglets (8 weeks old) and 3 pigs (35 weeks old) that were not vaccinated against FMDV. O-type FMDV (2 × 10² median tissue culture infectious dose) was inoculated into porcine PBMCs and the cells were incubated at 37.0℃ under 5% CO₂ for various time periods (0, 1, 3, 6, 12, 24, and 48 h). The total RNA was obtained from the FMDV-inoculated PBMCs after each time point, and the virus titer was investigated in these RNA samples. Furthermore, dynamics of mRNA expression of the six tested cytokines (interferon [IFN]-α, IFN-γ, interleukin [IL]-6, IL-8, IL-10, and tumor necrosis factor [TNF]-α) in FMDV-inoculated porcine PBMCs were evaluated by time-series analysis to determine the differences, if any, based on the age of the pigs. The PBMCs of piglets contained the highest quantity of FMDV mRNA at 6 hours post-inoculation (hpi), and the PBMCs of pigs had the highest quantity of FMDV mRNA at 3 hpi. The mean cycle threshold-value in the PBMCs steadily decreased after the peak time point in the piglets and pigs (6 and 3 hpi, respectively). The dynamics of mRNA expression of all cytokines except TNF-α showed age-dependent differences in FMDV-inoculated PBMCs. The mRNA expression of most cytokines was more pronounced in the piglets than in the pigs, implying that the immune response against FMDV showed an age-dependent difference in pigs. In conclusion, within 48 hpi, the 8-week-old piglets responded more rapidly and were more sensitive to FMDV infection than the 35-week-old pigs, which could be associated with the difference in the pathogenesis of FMDV infection among the pigs. These results provide valuable insights into the mechanisms underlying the age-dependent differences in immune response in pigs against FMDV infection.

• Journal of Community Nutrition
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• 제7권1호
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• pp.42-48
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• 2005

This study was done to investigate effects of antioxidant supplementation on phytohemagglutinin (PHA) -stimulated interleukin-2 (IL-2) production by peripheral blood mononuclear cells (PBMCs) in elderly women. This study was designed as a placebo-controlled, single-blinded, randomized intervention trial. Twenty four elderly women aged over 60 years, visitings social welfare center in Seoul were divided into 3 groups, placebo (n = 8), vitamin C supplemented (n = 8) , and vitamin E supplemented (n = 8) groups. Experimental groups were given either 1000mg of L-ascorbic acid or 400 ill of d- $\alpha$-tocopherol for 4 weeks. There was no significant difference in antioxidant vitamins intakes and their plasma levels among pre-intervention groups. Plasma vitamin C or E levels was significantly increased after vitamin C or E sup-plementations. The increases of plasma thiobarbituric acid-reactive substance (TBARS) levels in the placebo group were significantly higher than those of the supplemented 2 groups. There were no significant differences in the changes of plasma IL-2 level between pre- and post-intervention among the 3 groups. However there was a significant increase in PHA­stimulated IL-2 production by PBMCs after 4-week vitamin E or vitamin C supplementation. Particularly, vitamin E supplemented group showed a higher PHA-stimulated IL-2 production than vitamin C supplemented group. These results indicate that vitamin E or vitamin C supplementation might enhance mitogen-stimulated cytokine production by immune cells, which could be one of the factors to improve health status in the elderly.

• Asian-Australasian Journal of Animal Sciences
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• 제25권7호
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• pp.1038-1044
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• 2012

In the present study, the impact of Salmonella Typhimurium on cell-mediated immunity (CMI) was investigated in 5 week-old immuno divergent broiler lines selected for the high and low response to phytohemagglutinin-P. The immune response was assessed in peripheral-blood mononuclear cells (PBMCs) induced with Salmonella Typhimurium at different time intervals (0 h, 0.5 h, 2 h, 4 h, 6 h, 12 h and 24 h). The differential mRNA expression patterns of IFN-${\gamma}$, IL-2 and iNOS were evaluated by quantitative real time PCR. In-vitro production of nitric oxide (NO) was also estimated in the culture supernatant and correlated with iNOS mRNA expression. Present study showed higher production of NO in the high cell-mediated line (HCMI) as compared to the low cell-mediated line (LCMI) upon stimulation with Salmonella Typhimurium. Correspondingly, higher mRNA expression of iNOS and IFN-${\gamma}$ were observed in high response birds (HCMI); but IL-2 was down regulated in this line compared to the low response birds (LCMI). Significantly (p<0.05) higher expression of iNOS, IFN-${\gamma}$ and higher production of NO in high line indicated that the selection for PHA-P response might be employed for increasing the immune competence against Salmonella Typhimurium in chicken flocks.

• Molecules and Cells
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• 제45권10호
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• pp.738-748
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• 2022

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has posed a serious threat to global public health. A novel vaccine made from messenger RNA (mRNA) has been developed and approved for use at an unprecedented pace. However, an increased risk of myocarditis has been reported after BNT162b2 mRNA vaccination due to unknown causes. In this study, we used single-cell RNA sequencing and single-cell T cell receptor sequencing analyses of peripheral blood mononuclear cells (PBMCs) to describe, for the first time, changes in the peripheral immune landscape of a patient who underwent myocarditis after BNT162b2 vaccination. The greatest changes were observed in the transcriptomic profile of monocytes in terms of the number of differentially expressed genes. When compared to the transcriptome of PBMCs from vaccinated individuals without complications, increased expression levels of IL7R were detected in multiple cell clusters. Overall, results from this study can help advance research into the pathogenesis of BNT162b2-induced myocarditis.

• Asian Pacific Journal of Cancer Prevention
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• 제15권11호
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• pp.4455-4458
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• 2014

Breast cancer is a serious and potentially lethal multi-factor disease among 40-50 aged women in both developed and developing countries. Also, various studies have pointed to roles of neurotransmitters like serotonin in development of cancers, through action on various types of receptors. This study was conducted to evaluate serotonin receptor (5HT2AR and 5HT3AR) genes expression in peripheral blood mononuclear cells (PBMCs) of breast cancer patients in comparison with the healthy people and in the MCF7 cell line. Peripheral blood samples were obtained from 30 patients and 30 healthy individuals. Total RNA was extracted from PBMCs and MCF-7 cells. and 5HT2AR and 5HT3AR were detected by RT-PCR techniques. Finally, serotonin receptor gene expression variation in breast cancer patients and MCF-7 cells were determined by real time-PCR. This latter indicated significant promotion in expression of 5HT3AR and 5HT2AR in PBMCs in breast cancer patients but expression of 5HT2AR in the MCF-7 cell line was significantly decreased. In conclusion, after performing complimentary tests, determine of gene expression changes in serotonin receptors (5HT2AR and 5HT3AR) may be useful as a new approach in treatment of breast cancer based on use of antagonists.